LOCUS       sequence-291507-        6562 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6562)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046180
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2231..2248
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2249..2272
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2317..2335)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2686..3489
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3572..4554
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4632..4661
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4683..5270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5282..5309)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5333..5524)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5525..6316,6317..6385))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6148..6167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6386..6488)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6489..6520)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggttgggg cttgtgttct ttgcaattca cagacttcat taagatgtgg
      481 tgcttgcata cgtagaccat tcttatgttg taaatgctgt tacgaccatg tcatatcaac
      541 atcacataaa ttagtcttgt ctgttaatcc gtatgtttgc aatgctccag gttgtgatgt
      601 cacagatgtg actcaacttt acttaggagg tatgagctat tattgtaaat cacataaacc
      661 acccattagt tttccattgt gtgctaatgg acaagttttt ggtttatata aaaatacatg
      721 tgttggtagc gataatgtta ctgactttaa tgcaattgca acatgtgact ggacaaatgc
      781 tggtgattac attttagcta acacctgtac tgaaagactc aagctttttg cagcagaaac
      841 gctcaaagct actgaggaga catttaaact gtcttatggt attgctactg tacgtgaagt
      901 gctgtctgac agagaattac atctttcatg ggaagttggt aaacctagac caccacttaa
      961 ccgaaattat gtctttactg gttatcgtgt aactaaaaac agtaaagtac aaataggaga
     1021 gtacaccttt gaaaaaggtg actatggtga tgctgttgtt taccgaggta caacaactta
     1081 caaattaaat gttggtgatt attttgtgct gacatcacat acagtaatgc cattaagtgc
     1141 acctacacta gtgccacaag agcactatgt tagaattact ggcttatacc caacactcaa
     1201 tatctcagat gagttttcta gcaatgttgc aaattatcaa aaggttggta tgcaaaagta
     1261 ttctacactc cagggaccac ctggtactgg taagagtcat tttgctattg gcctagctct
     1321 ctactaccct tctgctcgca tagtgtatac agcttgctct catgccgctg ttgatgcact
     1381 atgtgagaag gcattaaaat atttgcctat agataaatgt agtagaatta tacctgcacg
     1441 tgctcgtgta gagtgttttg ataaattcaa agtgaattca acattagaac agtatgtctt
     1501 ttgtactgta aatgcattgc ctgagacgac agcagatata gttgtctttg atgaaatttc
     1561 aatggccaca aattatgatt tgagtgttgt caatgccaga ttacgtgcta agcactatgt
     1621 gtacattggc gaccctgctc aattacctgc accacgcaca ttgctaacta agggcacact
     1681 agaaccagaa tatttcaatt cagtgtgtag acttatgaaa actataggtc cagacatgtt
     1741 cctcggaact tgtcggcgtt gtcctgctga aattgttgac actgtgagtg ctttggttta
     1801 tgataataag cttaaagcac ataaagacaa atcagctcaa tgctttaaaa tgttttataa
     1861 gggtgttatc acgcatgatg tttcatctgc aattaacagg ccacaaatag gcgtggtaag
     1921 agaattcctt acacgtaacc ctgcttggag aaaagctgtc tttatttcac cttataattc
     1981 acagaatgct gtagcctcaa agattttggg actaccaact caaactgttg attcatcaca
     2041 gggctcagaa tatgactatg tcatattcac tcaaaccact gaaacagctc actcttgtaa
     2101 tgtaaacaga tttaatgttg ctattaccag agcaaaagta ggcatacttt gcataatgtc
     2161 tgatagagac ctttatgaca agttgcaatt tacaagtctt gaaattccac gtaggaatgt
     2221 ggcaacttta ctcgtgccga gaggttcttg gagccatccg cagtttgaaa aataaacagg
     2281 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     2341 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2401 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2461 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     2521 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2581 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2641 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     2701 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     2761 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     2821 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     2881 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     2941 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     3001 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     3061 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     3121 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     3181 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     3241 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     3301 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     3361 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     3421 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     3481 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     3541 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     3601 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     3661 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     3721 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3781 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     3841 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     3901 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3961 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     4021 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4081 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     4141 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4201 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4261 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4321 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4381 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4441 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     4501 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     4561 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     4621 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     4801 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5161 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5221 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5281 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     5341 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5401 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5461 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5521 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     5641 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5701 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     5761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5821 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5941 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     6001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     6061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6181 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     6241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6361 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     6421 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     6481 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     6541 atgatttaaa tggtcagtta ga
//
